An episomal CRISPR/Cas9 system to derive vector-free gene modified mammalian cells
Dear Editor,CRISPR and CRISPR-associated (Cas) proteins play their adaptive immunity role in degrading foreign nucleic acids in both bacteria and archaea.CRISPR/Cas has proved efficient in modifying mammalian genomes (Cong et al.,2013;Mali et al.,2013) and various delivery methods for Cas9:gRNA complex have been established.Among different delivery means,in vitro transcribed Cas9 mRNA/gRNA,purified Cas9 protein,and adeno-associated virus (AAV) driven seem promising toward clinical applications (Kouranova et al.,2016;Lin et al.,2014;Yin et al.,2016).Still,all the fore-mentioned methods for delivering CRISPR/Cas9 have certain shortcomings./n vitro transcription and protein purification are complicated and inconvenient for use,while AAV mediated system suffers from the side effects of immune response and insertional mutagenesis (Donsante et al.,2007;Li et al.,2011).
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This work was supported by the National Natural Science Foundation of China Grant No.31271598 and The Project for Extramural Scientists of State Key Laboratory of Agrobiotechnology 2015SKLAB6-15.
2016-11-08（万方平台首次上网日期，不代表论文的发表时间）
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